Expression of the genes encoding basic fibroblastgrowth factor and its receptorin human granulosa cells

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Abstract

The role of basic fibroblast growth factor (bFGF) in granulosa cell ontogeny has been previously demonstrated. In this study we evaluated the possible intraovarian origin of bFGF. Human granulosa cells were maintained in primary culture and their cytoplasmic extract was purified by affinity chromatography on a heparin-sepharose column. The column was then eluted with 10 mM Tris-HCl containing increasing concentrations of NaCl. The Chromatographie fractions were tested in a bioassay using bovine adrenal capillary endothelial cells (ACE) as targets. A peak of mitogenic activity was detected in the fraction eluted with the highest salt concentration. This Chromatographie profile is similar to that of bFGF. The in situ synthesis of this bFGF-like protein was then demonstrated by reverse transcription-polymerase chain reaction. Using oligonucleotide primers specific for the bFGF gene, a single major band of DNA, corresponding to the expected size, was amplified. The identity of this fragment with the bFGF corresponding sequence was further demonstrated by restriction enzyme analysis. Moreover, RT-PCR was also employed to amplify a DNA band specific for the bFGF receptor gene. These data indicate that human granulosa cells are able to synthesize both bFGF and its receptor and, thus, bFGF might participate to the autocrine mechanisms regulating their growth and differentiated functions.

Original languageEnglish
JournalMolecular and Cellular Endocrinology
Volume96
Issue number1-2
DOIs
Publication statusPublished - 1993

Fingerprint

Gene encoding
Granulosa Cells
Fibroblast Growth Factor 2
Gene Expression
Fibroblast Growth Factor Receptors
Genes
Affinity chromatography
Polymerase Chain Reaction
Restriction Mapping
DNA Primers
Bioassay
Polymerase chain reaction
DNA
Endothelial cells
Transcription
Affinity Chromatography
Biological Assay
Reverse Transcription
Endothelial Cells
Salts

Keywords

  • Basic fibroblast growth factor
  • Granulosa cell
  • Polymerase chain reaction

ASJC Scopus subject areas

  • Endocrinology
  • Endocrinology, Diabetes and Metabolism

Cite this

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title = "Expression of the genes encoding basic fibroblastgrowth factor and its receptorin human granulosa cells",
abstract = "The role of basic fibroblast growth factor (bFGF) in granulosa cell ontogeny has been previously demonstrated. In this study we evaluated the possible intraovarian origin of bFGF. Human granulosa cells were maintained in primary culture and their cytoplasmic extract was purified by affinity chromatography on a heparin-sepharose column. The column was then eluted with 10 mM Tris-HCl containing increasing concentrations of NaCl. The Chromatographie fractions were tested in a bioassay using bovine adrenal capillary endothelial cells (ACE) as targets. A peak of mitogenic activity was detected in the fraction eluted with the highest salt concentration. This Chromatographie profile is similar to that of bFGF. The in situ synthesis of this bFGF-like protein was then demonstrated by reverse transcription-polymerase chain reaction. Using oligonucleotide primers specific for the bFGF gene, a single major band of DNA, corresponding to the expected size, was amplified. The identity of this fragment with the bFGF corresponding sequence was further demonstrated by restriction enzyme analysis. Moreover, RT-PCR was also employed to amplify a DNA band specific for the bFGF receptor gene. These data indicate that human granulosa cells are able to synthesize both bFGF and its receptor and, thus, bFGF might participate to the autocrine mechanisms regulating their growth and differentiated functions.",
keywords = "Basic fibroblast growth factor, Granulosa cell, Polymerase chain reaction",
author = "{Di Blasio}, {Anna Maria} and Paola Vigano and Laura Cremonesi and Cristiana Carniti and Maurizio Ferrari and Augusto Ferrari",
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T1 - Expression of the genes encoding basic fibroblastgrowth factor and its receptorin human granulosa cells

AU - Di Blasio, Anna Maria

AU - Vigano, Paola

AU - Cremonesi, Laura

AU - Carniti, Cristiana

AU - Ferrari, Maurizio

AU - Ferrari, Augusto

PY - 1993

Y1 - 1993

N2 - The role of basic fibroblast growth factor (bFGF) in granulosa cell ontogeny has been previously demonstrated. In this study we evaluated the possible intraovarian origin of bFGF. Human granulosa cells were maintained in primary culture and their cytoplasmic extract was purified by affinity chromatography on a heparin-sepharose column. The column was then eluted with 10 mM Tris-HCl containing increasing concentrations of NaCl. The Chromatographie fractions were tested in a bioassay using bovine adrenal capillary endothelial cells (ACE) as targets. A peak of mitogenic activity was detected in the fraction eluted with the highest salt concentration. This Chromatographie profile is similar to that of bFGF. The in situ synthesis of this bFGF-like protein was then demonstrated by reverse transcription-polymerase chain reaction. Using oligonucleotide primers specific for the bFGF gene, a single major band of DNA, corresponding to the expected size, was amplified. The identity of this fragment with the bFGF corresponding sequence was further demonstrated by restriction enzyme analysis. Moreover, RT-PCR was also employed to amplify a DNA band specific for the bFGF receptor gene. These data indicate that human granulosa cells are able to synthesize both bFGF and its receptor and, thus, bFGF might participate to the autocrine mechanisms regulating their growth and differentiated functions.

AB - The role of basic fibroblast growth factor (bFGF) in granulosa cell ontogeny has been previously demonstrated. In this study we evaluated the possible intraovarian origin of bFGF. Human granulosa cells were maintained in primary culture and their cytoplasmic extract was purified by affinity chromatography on a heparin-sepharose column. The column was then eluted with 10 mM Tris-HCl containing increasing concentrations of NaCl. The Chromatographie fractions were tested in a bioassay using bovine adrenal capillary endothelial cells (ACE) as targets. A peak of mitogenic activity was detected in the fraction eluted with the highest salt concentration. This Chromatographie profile is similar to that of bFGF. The in situ synthesis of this bFGF-like protein was then demonstrated by reverse transcription-polymerase chain reaction. Using oligonucleotide primers specific for the bFGF gene, a single major band of DNA, corresponding to the expected size, was amplified. The identity of this fragment with the bFGF corresponding sequence was further demonstrated by restriction enzyme analysis. Moreover, RT-PCR was also employed to amplify a DNA band specific for the bFGF receptor gene. These data indicate that human granulosa cells are able to synthesize both bFGF and its receptor and, thus, bFGF might participate to the autocrine mechanisms regulating their growth and differentiated functions.

KW - Basic fibroblast growth factor

KW - Granulosa cell

KW - Polymerase chain reaction

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